Bisulfite Sequencing of RNA for Transcriptome-Wide Detection of 5-Methylcytosine

Methods Mol Biol. 2019:1870:1-21. doi: 10.1007/978-1-4939-8808-2_1.

Abstract

A powerful method to determine the methylation status of specific cytosine residues within RNA is bisulfite sequencing. In combination with high-throughput sequencing methods cytosine methylation can be determined at nucleotide resolution on a transcriptome-wide level. Nevertheless, several critical aspects need to be considered before starting such a project. Below we describe a detailed step-by-step protocol for planning and performing a transcriptome-wide bisulfite sequencing experiment and subsequent data analysis to determine methyl-cytosine in poly(A)RNA from cells and tissues.

Keywords: 5-Methylcytosine; Bisulfite sequencing; High throughput; Next generation sequencing; RNA methylation; Transcriptome; meRanTK.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5-Methylcytosine* / metabolism
  • Gene Expression Profiling
  • High-Throughput Nucleotide Sequencing
  • Methylation
  • RNA / chemistry
  • RNA / genetics*
  • RNA Folding
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics
  • Sequence Analysis, DNA
  • Transcriptome*

Substances

  • RNA, Messenger
  • RNA
  • 5-Methylcytosine